Glycerol was transglycosylated by cyclodextrin glucanotransferases using starch as a donor substrate. Among the enzymes tested, those from Geobacillus stearothermophilus and Thermoanaerobacter sp. were suitable for the transglycosylation. Several products were isolated and their structures were elucidated. They were composed of glucose and a series of a-1,4-linked maltooligosyl residues bound with glycerol. O-alpha-D-Glucosyl-(1-->1)-glycerol and O-alpha-D-glucosyl-(1-->2)-glycerol were identified as the major and minor components of the smallest transfer products, respectively. O-alpha-D-Glucosyl-(1-->4)-O-alpha-D-glucosyl-(1-->1)-glycerol was also identified as a main dimer product. Reducing sugars were produced in extremely low amounts. The optimum temperatures for the transglycosylation by G. stearothermophilus and Thermoanaerobacter enzymes were approximately 60 degrees C and 80 degrees C, respectively. The reaction of 30% (w/v) glycerol and 20% (w/v) soluble starch was optimum for efficient transglycosylation. Maltosyl and maltotriosyl glycerols inhibited porcine pancreas a-amylase significantly, whereas the monomer, glucosyl glycerol, exhibited much weaker inhibition.